鞣花酸通过调控miR-1254表达抑制口腔鳞癌HSC3细胞增殖、迁移和侵袭的机制

Ellagic acid inhibits the proliferation,migration and invasion of the oral squamous cell carcinoma cell line HSC3 by regulating miR-1254

目的 探讨鞣花酸对口腔鳞癌HSC3细胞增殖、迁移和侵袭的影响及可能机制。方法 体外培养HSC3细胞,用不同剂量(2、4、8μg/ml)鞣花酸干预24 h后,CCK-8法和克隆形成实验检测细胞增殖、划痕实验和Transwell分别检测细胞迁移和侵袭,蛋白质印迹法检测细胞中E-cadherin和N-cadherin蛋白表达,qRT-PCR法检测细胞中miR-1254表达。收集39例口腔鳞癌患者癌组织和癌旁组织,RT-PCR法检测组织中miR-1254表达。转染miR-1254模拟物至HSC3细胞,或转染miR-1254抑制剂至HSC3细胞后再用8μg/ml鞣花酸干预24 h,然后采用上述相同方法检测细胞增殖、迁移和侵袭。结果 与对照组比较,不同剂量鞣花酸提高了HSC3细胞增殖抑制率和E-cadherin蛋白表达(P<0.05),降低了细胞克隆形成数、划痕愈合率、侵袭数及N-cadherin蛋白表达(P<0.05)。同时,与对照组比较,不同剂量鞣花酸促进了HSC3细胞中miR-1254表达(P<0.05)。口腔鳞癌组织中miR-1254表达低于癌旁组织(P<0.05)。过表达miR-1254后,HSC3细胞增殖抑制率和E-cadherin蛋白表达升高(P<0.05),细胞克隆形成数、划痕愈合率、侵袭数及N-cadherin蛋白表达降低(P<0.05)。过表达miR-1254逆转了鞣花酸对HSC3细胞增殖、迁移和侵袭的影响。结论 鞣花酸可能通过上调miR-1254抑制口腔鳞癌HSC3细胞增殖、迁移和侵袭。

Objective To investigate the effect of ellagic acid on the proliferation,migration and invasion of the oral squamous cell carcinoma cell line HSC3 and its possible mechanism.Methods HSC3 cells were cultured in vitro and treated with different doses(2,4,8μg/ml)of ellagic acid for 24h.Cell proliferation was detected by CCK-8 assay and clone formation test.Cell migration and invasion were detected by scratch test and Transwell assay,respectively.Western blotting was used to detect the protein expressions of E-cadherin and N-cadherin,and qRT-PCR was used to detect the expression of miR-1254.Moreover,relative levels of miR-1254 in 39 pairs of oral squamous cell carcinoma specimens and paracancerous specimens were measured by qRT-PCR.Cell proliferation,migration and invasion in HSC3 cells transfected with miR-1254 mimics or mimic control,followed by the treatment of 8μg/ml ellagic acid for 24h.Results Treatment of different doses of ellagic acid increased the proliferation inhibition rate and the protein expression of E-cadherin in HSC3 cells(P<0.05),but reduced the number of cell clones,scratch healing rate,the number of invasive cells,and the protein expression of N-cadherin(P<0.05),which also upregulated miR-1254 in HSC3 cells.The expression of miR-1254 in oral squamous cell carcinoma specimens was significantly lower than that in paracancerous specimens(0.27±0.05 vs 1.00±0.13,P<0.05).After overexpression of miR-1254,the proliferation inhibition rate and the protein expression of E-cadherin in HSC3 cells were significantly increased(P<0.05),but the number of clones,scratch healing rate,the number of invasive cells,and the protein expression of N-cadherin were significantly reduced(P<0.05).Overexpression of miR-1254 reversed the effect of ellagic acid on the proliferation,migration and invasion of HSC3 cells.Conclusion Ellagic acid may inhibit the proliferation,migration and invasion of the oral squamous cell carcinoma cell line HSC3 by upregulating miR-1254.